TY - JOUR
T1 - An assay to image neuronal microtubule dynamics in mice
AU - Kleele, Tatjana
AU - Marinković, Petar
AU - Williams, Philip R.
AU - Stern, Sina
AU - Weigand, Emily E.
AU - Engerer, Peter
AU - Naumann, Ronald
AU - Hartmann, Jana
AU - Karl, Rosa M.
AU - Bradke, Frank
AU - Bishop, Derron
AU - Herms, Jochen
AU - Konnerth, Arthur
AU - Kerschensteiner, Martin
AU - Godinho, Leanne
AU - Misgeld, Thomas
N1 - Publisher Copyright:
© 2014 Macmillan Publishers Limited. All rights reserved.
PY - 2014
Y1 - 2014
N2 - Microtubule dynamics in neurons play critical roles in physiology, injury and disease and determine microtubule orientation, the cell biological correlate of neurite polarization. Several microtubule binding proteins, including end-binding protein 3 (EB3), specifically bind to the growing plus tip of microtubules. In the past, fluorescently tagged end-binding proteins have revealed microtubule dynamics in vitro and in non-mammalian model organisms. Here, we devise an imaging assay based on transgenic mice expressing yellow fluorescent protein-tagged EB3 to study microtubules in intact mammalian neurites. Our approach allows measurement of microtubule dynamics in vivo and ex vivo in peripheral nervous system and central nervous system neurites under physiological conditions and after exposure to microtubule-modifying drugs. We find an increase in dynamic microtubules after injury and in neurodegenerative disease states, before axons show morphological indications of degeneration or regrowth. Thus increased microtubule dynamics might serve as a general indicator of neurite remodelling in health and disease.
AB - Microtubule dynamics in neurons play critical roles in physiology, injury and disease and determine microtubule orientation, the cell biological correlate of neurite polarization. Several microtubule binding proteins, including end-binding protein 3 (EB3), specifically bind to the growing plus tip of microtubules. In the past, fluorescently tagged end-binding proteins have revealed microtubule dynamics in vitro and in non-mammalian model organisms. Here, we devise an imaging assay based on transgenic mice expressing yellow fluorescent protein-tagged EB3 to study microtubules in intact mammalian neurites. Our approach allows measurement of microtubule dynamics in vivo and ex vivo in peripheral nervous system and central nervous system neurites under physiological conditions and after exposure to microtubule-modifying drugs. We find an increase in dynamic microtubules after injury and in neurodegenerative disease states, before axons show morphological indications of degeneration or regrowth. Thus increased microtubule dynamics might serve as a general indicator of neurite remodelling in health and disease.
UR - http://www.scopus.com/inward/record.url?scp=84908256346&partnerID=8YFLogxK
U2 - 10.1038/ncomms5827
DO - 10.1038/ncomms5827
M3 - Article
C2 - 25219969
AN - SCOPUS:84908256346
SN - 2041-1723
VL - 5
JO - Nature communications
JF - Nature communications
M1 - 4827
ER -